Validation of a cell-based assay to differentiate between the cytotoxic effects of elapid snake venoms

Kalam, Yasmean; Isbister, Geoffrey K.; Mirtschin, Peter; Hodgson, Wayne Clarence; Konstantakopoulos, Nicki

doi:10.1016/j.vascn.2010.09.001

Cited by 25 publications

(25 citation statements)

References 13 publications

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“…Our interpretation of the results suggests that the LD 50 value is approximately ten times greater than the IC 50 value for crude venom from Macrovipera lebetina lebetina. Our results agree with those from other authors (40)(41)(42). Another important point is that the venom enhances the cytotoxic response of L929 cells when administered for longer periods (48 hours) with an IC 50 value of 0.62 ± 0.18 µg/ mL.…”

Section: Discussionsupporting

confidence: 93%

“…Venom-antivenom neutralizing ability. On the other hand, according to our results and previous studies, the cell-based assay can be useful for examining cytotoxicity levels and the ability of antivenom to neutralize snake venom (18,19,41). However, in vitro tests are not currently good enough to replace all animal tests as the in vitro methodology needs further optimization.…”

Section: Discussionmentioning

confidence: 76%

“…Currently, researchers suggest that cell-based assay to examine venom cytotoxicity is an alternative to animal testing (18,19,(40)(41)(42). Our in vitro results show that crude venom from Macrovipera lebetina lebetina is highly cytotoxic for cultured fibroblasts causing decreased viability, the disappearance of normal morphological characteristics, rounding up, detachment and death at the highest concentration.…”

Section: Discussionmentioning

confidence: 77%

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Determination of in vivo toxicity and in vitro cytotoxicity of venom from the Cypriot blunt-nosed viper Macrovipera lebetina lebetina and antivenom production

Nalbantsoy¹,

Karabay-Yavasoglu²,

Sayim³

et al. 2012

J. Venom. Anim. Toxins incl. Trop. Dis

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Abstract:The venomous Levantine viper, Macrovipera lebetina lebetina is endemic to Cyprus. The objective of this study was to investigate in vitro cytotoxicity, in vivo lethality, and antivenom production followed by a re-immunization schedule in mice against Macrovipera lebetina lebetina venom. The LD 50 value was estimated as 7.58mg/kg within 24 hours by different venom doses administrated intraperitoneally in mice. Freund's complete and incomplete adjuvants were used for first and second immunization of mice in antivenom production. A cell-based assay was performed to determine the effects of Macrovipera lebetina lebetina venom and antivenom neutralizing potency on L929 cell viability. The snake venom toxicity and cytotoxicity were examined and comparison of results showed good correlation, the LD 50 value was tenfold higher than the IC 50 value. The IC 50 value was 0.62 ± 0.18 µg/mL after 48 hours treatment while the calculated value was 1.62 ± 0.25 µg/mL for the culture media totally refreshed after two hours treatment with venom. The in vitro efficacy of antivenom against Macrovipera lebetina lebetina venom was found to be low. This is the first report that describes the in vivo and in vitro toxic effects of Macrovipera lebetina lebetina venom and antivenom production against this species.

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Section: Discussionsupporting

confidence: 93%

Section: Discussionmentioning

confidence: 76%

Section: Discussionmentioning

confidence: 77%

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Determination of in vivo toxicity and in vitro cytotoxicity of venom from the Cypriot blunt-nosed viper Macrovipera lebetina lebetina and antivenom production

Nalbantsoy¹,

Karabay-Yavasoglu²,

Sayim³

et al. 2012

J. Venom. Anim. Toxins incl. Trop. Dis

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“…To date, there are very few published works that have studied and compared the cytotoxicity of venoms from different snake species in one study and none assessing snakes found in Malaysia (6,7). The most useful information found by this assay is the number of viable cells at the end of the snake venom treatment.…”

Section: Discussionmentioning

confidence: 99%

Cytotoxicity of Southeast Asian snake venoms

Jamunaa¹,

Vejayan²,

Halijah³

et al. 2012

J. Venom. Anim. Toxins incl. Trop. Dis

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Cytotoxicity of venoms from eleven medically important snakes found in Southeast Asia (Naja kaouthia, Naja siamensis, Naja sumatrana, Ophiophagus hannah, Bungarus candidus, Bungarus fasciatus, Enhydrina schistosa, Calloselasma rhodostoma, Trimeresurus purpureomaculatus and Tropidolaemus sumatranus) was determined, based on the MTS cytotoxicity assay, which determines the survival of viable cells in monolayer MDCK and Vero cell cultures upon exposure to the snake venoms. Snake venom toxicity was expressed as the venom dose that killed 50% of the cells (CTC 50 ) under the assay conditions. Venoms of C. rhodostoma (2.6 µg/mL, 1.4 µg/mL) and O. hannah were the most cytotoxic (3.8 µg/mL, 1.7 µg/mL) whereas N. siamensis venom showed the least cytotoxicity (51.9 µg/mL, 45.7 µg/mL) against Vero and MDCK cells, respectively. All the viper venoms showed higher cytotoxic potency towards both Vero and MDCK cell lines, in comparison to krait and cobra venoms. E. schistosa did not cause cytotoxicity towards MDCK or Vero cells at the tested concentrations. The cytotoxicity correlates well with the known differences in the composition of venoms from cobras, kraits, vipers and sea snakes.

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“…Recently, a MTS based cell cytotoxicity assay is being widely used as an alternative model for testing the cytotoxic effect of venoms as well as the efficacy of antivenoms with the advantage of avoiding the use of experimental animals [6,12,13]. In this study, a rat skeletal muscle cell line, L6, was used to examine the cytotoxic activity of Naja nubiae and Echis ocellatus venoms.…”

Section: Introductionmentioning

confidence: 99%

Cytotoxiciy of Naja nubiae (Serpentes: Elapidae) and Echis ocellatus (Serpentes: Viperidae) Venoms from Sudan

2015

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In Sudan, as in many African countries, no local specific antivenom is manufactured resulting in snake bite victims being treated by antivenoms imported from abroad. In the present work we measured the cytotoxic effect of the recently described spitting cobra (Naja nubiae) and the carpet viper (Echis ocellatus) snake venoms using a cell based assay. We also investigated the efficacy of four antivenoms CSL (Australia), SAIMR (South Africa), snake venom antiserum (India), and EchiTAb-Plus-ICP (Cost Rica) to neutralize the cytotoxic effect of the two venoms. The venoms resulted in a remarkable inhibition of cell viability with N. nubiae being more cytotoxic than E. ocellatus. The four antivenoms studied were effective in neutralizing N. nubiae cytotoxicity. However, only partial efficacy in neutralizing the cytotoxic effect of E. ocellatus was achieved using CSL (Australia) and SVA (India) antivenoms. Based on the cross neutralization by the four antivenoms, the Sudanese N. nubiae venom most likely has homologous epitopes with similar snakes from Australia, South Africa, India, and Cost Rica, while E. ocellatus venom from Sudan shares little homology with similar snakes from other countries.

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Validation of a cell-based assay to differentiate between the cytotoxic effects of elapid snake venoms

Cited by 25 publications

References 13 publications

Determination of in vivo toxicity and in vitro cytotoxicity of venom from the Cypriot blunt-nosed viper Macrovipera lebetina lebetina and antivenom production

Determination of in vivo toxicity and in vitro cytotoxicity of venom from the Cypriot blunt-nosed viper Macrovipera lebetina lebetina and antivenom production

Cytotoxicity of Southeast Asian snake venoms

Cytotoxiciy of Naja nubiae (Serpentes: Elapidae) and Echis ocellatus (Serpentes: Viperidae) Venoms from Sudan

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